title: Conditioned medium from amniotic membrane-derived cells prevents lung fibrosis and preserves blood gas exchanges in bleomycin-injured mice-specificity of the effects and insights into possible mechanisms.
abstract: OBJECTIVE
  We recently demonstrated that injection of conditioned medium (CM) generated from cells of the mesenchymal region of human amniotic membrane (AMTCs) reduces bleomycin-induced lung fibrosis in mice, suggesting a crucial role of paracrine factor(s) secreted by AMTCs in these beneficial effects. We further investigated this hypothesis, the mechanisms involved, the effects on some lung functional parameters and whether AMTC-secreted effector(s) are specific to these cells and not produced by other cell types, extending the time of analysis up to 28 days after treatment.
  
  
  METHODS
  Bleomycin-challenged mice were either treated with AMTC-CM or CM generated from human skin fibroblasts, human peripheral blood mononuclear cells or Jurkat cells, or were left untreated. Mouse lungs were analyzed for content of pro-inflammatory and pro-fibrotic molecules, presence of lymphocytes and macrophages and for fibrosis level (through histological semi-quantitative evaluation and quantitative measurement of collagen content). Arterial blood gas analysis was also performed.
  
  
  RESULTS
  Up to 28 days after delivery, AMTC-CM-treated mice developed reduced lung fibrosis with respect to mice treated with other CM types. AMTC-CM-treated mice had comparatively better preservation of blood gas parameters and showed lower lung content of interleukin-6, tumor necrosis factor-α, macrophage inflammatory protein-1α, monocyte chemoattractant protein-1 and transforming growth factor-β associated with reduced lung macrophage levels.
  
  
  CONCLUSIONS
  AMTC-CM prevents lung fibrosis in bleomycin-challenged mice, improving survival and preserving lung functional parameters such as blood gas exchanges. The specificity of AMTC-CM action was indicated by the absence of fibrosis reduction when other CM types were used. Finally, we provide some insights into the possible mechanisms underlying AMTC-CM-mediated control of fibrosis.
authors.affiliation: Centro di Ricerca E. Menni, Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Cargnoni
authors.affiliation: Centro di Ricerca E. Menni, Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy.
authors.forename: Ester Cotti
authors.initials: EC
authors.lastname: Piccinelli
authors.affiliation: Centro di Ricerca E. Menni, Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy; School of Veterinary Science, University of Liverpool, Neston, United Kingdom.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Ressel
authors.affiliation: Centro di Ricerca E. Menni, Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Rossi
authors.affiliation: Centro di Ricerca E. Menni, Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Magatti
authors.affiliation: Dipartimento di Scienze Agrarie e Ambientali, Università di Milano, Milano, Italy.
authors.forename: Ivan
authors.initials: I
authors.lastname: Toschi
authors.affiliation: Dipartimento di Scienze Agrarie e Ambientali, Università di Milano, Milano, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Cesari
authors.affiliation: Dipartimento di Scienze Veterinarie e Sanità Pubblica, Università di Milano, Milano, Italy.
authors.forename: Mariangela
authors.initials: M
authors.lastname: Albertini
authors.affiliation: Dipartimento di Scienze Veterinarie e Sanità Pubblica, Università di Milano, Milano, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Mazzola
authors.affiliation: Centro di Ricerca E. Menni, Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy. Electronic address: ornella.parolini@tin.it.
authors.forename: Ornella
authors.initials: O
authors.lastname: Parolini
mesh_terms: D000650:Amnion / Q000166:cytology / Q000378:metabolism; D000818:Animals; D001761:Bleomycin / Q000633:toxicity; D001784:Blood Gas Analysis; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D004195:Disease Models, Animal; D006801:Humans; D008168:Lung / Q000187:drug effects* / Q000473:pathology; D051379:Mice; D011658:Pulmonary Fibrosis / Q000139:chemically induced / Q000188:drug therapy* / Q000473:pathology
chemical_list: D017077:Culture Media, Conditioned; D001761:Bleomycin
keywords: amniotic membrane-derived cells; amniotic mesenchymal tissue cells; conditioned medium; human term placenta; lung fibrosis; mesenchymal stromal cell